4.8 Article

Dual-Readout Sandwich Immunoassay for Device-Free and Highly Sensitive Anthrax Biomarker Detection

Journal

ANALYTICAL CHEMISTRY
Volume 92, Issue 11, Pages 7845-7851

Publisher

AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.0c01090

Keywords

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Funding

  1. Air Force Research Laboratory [FA8650-15-2-5518]
  2. Office of the Director of National Intelligence, Intelligence Advanced Research Projects Activity, via Federal Bureau of Investigation [DJF-15-1200-K-0001730]
  3. Air Force Office of Scientific Research Awards [FA9550-18-1-0493, FA9550-16-1-0150]
  4. NTU-NU Institute for NanoMedicine
  5. National Cancer Institute of the National Institutes of Health [U54CA199091, U54CA151880]
  6. Cellular and Molecular Basis of Disease Training Program - National Institutes of Health, Institute of General Medical Sciences Award [T32GM008061]
  7. Chemistry of Life Processes Training Program - National Institute of General Medical Sciences Award [T32GM105538]
  8. Nanyang Technological University, Singapore

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We report a dual-readout, AuNP-based sandwich immuno-assay for the device-free colorimetric and sensitive scanometric detection of disease biomarkers. An AuNP-antibody conjugate serves as a signal transduction and amplification agent by promoting the reduction and deposition of either platinum or gold onto its surface, generating corresponding colorimetric or light scattering (scanometric) signals, respectively. We apply the Pt-based colorimetric readout of this assay to the discovery of a novel monoclonal antibody (mAb) sandwich pair for the detection of an anthrax protective antigen (PA(83)). The identified antibody pair detects PA(83) down to 1 nM in phosphate-buffered saline and 5 nM in human serum, which are physiologically relevant concentrations. Reducing gold rather than platinum onto the mAb-AuNP sandwich enables scanometric detection of subpicomolar PA(83) concentrations, over 3 orders of magnitude more sensitive than the colorimetric readout.

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